Efectos del cannabidiol en un nuevo modelo animal de trastorno de estrés postramático

En esta tesis doctoral se ha desarrollado un nuevo modelo animal de trastorno de estrés postraumático (TEPT) que induce cambios pronunciados sobre el condicionamiento al miedo y la ansiedad, que se mantienen hasta 9 semanas después de haber finalizado la fase de inducción (Artículo 1). Estas modificaciones se acompañan de alteraciones en la expresión génica de la hormona liberadora de corticotropina (CRF), receptores cannabinoides 1 (CNR1) y 2 (CNR2) y transportador de serotonina (5HTT) en distintos núcleos cerebrales. Debido a la larga duración de las características conductuales y de cambios funcionales cerebrales, el modelo ha permitido la evaluación de tratamientos farmacológicos de varias semanas de duración. Se ha observado que la administración de cannabidiol (CBD) o sertralina (STR) regula las alteraciones producidas por el modelo, si bien se consigue un efecto sinérgico o aditivo con la combinación de ambos fármacos (Artículo 1). Por otra parte, la larga duración de las alteraciones inducidas por el modelo ha permitido su combinación con los procedimientos experimentales del consumo voluntario (CV) y autoadministración oral de etanol (AAO). Los roedores expuestos al modelo animal de TEPT presentan una mayor vulnerabilidad por los efectos reforzantes y motivacionales del alcohol, simulando de esta manera el consumo problemático de alcohol en pacientes con TEPT. Estas conductas se acompañan de alteraciones en la expresión génica de la tirosina hidroxilasa (TH), receptor mu-opioide (Opmr1) y 5HTT en distintos núcleos cerebrales, indicando la implicación de los sistemas dopaminérgico, opioide y serotoninérgico en el desarrollo de la conducta. La administración de STR y naltrexona (NTX) reduce el consumo y la motivación por obtener alcohol en ratones expuestos al modelo animal de TEPT y regula los cambios en la expresión génica de las distintas dianas evaluadas (Artículo 2). En conjunto, los datos obtenidos en la presente tesis doctoral ponen de manifiesto que: i) el nuevo modelo animal crónico de TEPT desarrollado produce alteraciones en la respuesta emocional que se mantienen a largo plazo, incluyendo un mayor rasgo de ansiedad, problemas en la extinción de memorias aversivas (miedo), y una elevada vulnerabilidad por la acción reforzante y motivacional del alcohol, así como cambios significativos en la expresión génica de distintas dianas, ii) la combinación farmacológica de CBD y STR puede resultar un tratamiento eficaz en pacientes con TEPT, iii) la combinación de la STR con una dosis subefectiva de NTX podría ser una herramienta a tener en cuenta en el tratamiento del trastorno dual TEPT y trastornos por uso de alcohol (TUA)

Pairing Binge Drinking and a High-Fat Diet in Adolescence Modulates the Inflammatory Effects of Subsequent Alcohol Consumption in Mice

Alcohol binge drinking (BD) and poor nutritional habits are two frequent behaviors among many adolescents that alter gut microbiota in a pro-inflammatory direction. Dysbiotic changes in the gut microbiome are observed after alcohol and high-fat diet (HFD) consumption, even before obesity onset. In this study, we investigate the neuroinflammatory response of adolescent BD when combined with a continuous or intermittent HFD and its effects on adult ethanol consumption by using a self-administration (SA) paradigm in mice. The inflammatory biomarkers IL-6 and CX3CL1 were measured in the striatum 24 h after BD, 3 weeks later and after the ethanol (EtOH) SA. Adolescent BD increased alcohol consumption in the oral SA and caused a greater motivation to seek the substance. Likewise, mice with intermittent access to HFD exhibited higher EtOH consumption, while the opposite effect was found in mice with continuous HFD access. Biochemical analyses showed that after BD and three weeks later, striatal levels of IL-6 and CX3CL1 were increased. In addition, in saline-treated mice, CX3CL1 was increased after continuous access to HFD. After oral SA procedure, striatal IL-6 was increased only in animals exposed to BD and HFD. In addition, striatal CX3CL1 levels were increased in all BD- and HFD-exposed groups. Overall, our findings show that adolescent BD and intermittent HFD increase adult alcohol intake and point to neuroinflammation as an important mechanism modulating this interaction.021 by the authors. Licensee MDPI, Basel, Switzerland

Biomarkers of the Endocannabinoid System in Substance Use Disorders

Despite substance use disorders (SUD) being one of the leading causes of disability and mortality globally, available therapeutic approaches remain ineffective. The difficulty in accurately characterizing the neurobiological mechanisms involved with a purely qualitative diagnosis is an obstacle to improving the classification and treatment of SUD. In this regard, identifying central and peripheral biomarkers is essential to diagnosing the severity of drug dependence, monitoring therapeutic efficacy, predicting treatment response, and enhancing the development of safer and more effective pharmacological tools. In recent years, the crucial role that the endocannabinoid system (ECS) plays in regulating the reinforcing and motivational properties of drugs of abuse has been described. This has led to studies characterizing ECS alterations after exposure to various substances to identify biomarkers with potential diagnostic, prognostic, or therapeutic utility. This review aims to compile the primary evidence available from rodent and clinical studies on how the ECS components are modified in the context of different substance-related disorders, gathering data from genetic, molecular, functional, and neuroimaging experimental approaches. Finally, this report concludes that additional translational research is needed to further characterize the modifications of the ECS in the context of SUD, and their potential usefulness in the necessary search for biomarkers

Palaeoenvironmental and chronological context of hominin occupations of the Armenian Highlands during MIS 3:Evidence from Ararat-1 cave

Archaeological and palaeoenvironmental evidence from the Armenian Highlands and wider southern Caucasus region emphasises the significance of Marine Oxygen Isotope Stage 3 (c. 57–29 ka) as a crucial period for understanding hominin behaviours amidst environmental fluctuations. Ararat-1 cave, situated in the Ararat Depression, Republic of Armenia, presents potential for resolving emerging key debates regarding hominin land use adaptations during this interval, due to its well-preserved lithic artefacts and faunal assemblages. We present the first results of combined sedimentological, geochronological (luminescence and radiocarbon), archaeological and palaeoecological (macrofauna, microfauna and microcharcoal) study of the Ararat-1 sequence. We demonstrate sediment accumulation occurred between 52 and 35 ka and was caused by a combination of aeolian activity, cave rockfall and water action. Whilst the upper strata of the Ararat-1 sequence experienced postdepositional disturbance due to faunal and anthropogenic processes, the lower strata remain relatively undisturbed. We suggest that during a stable period within MIS 3, Ararat-1 was inhabited by Middle Palaeolithic hominins amidst a mosaic of semi-arid shrub, grassland, and temperate woodland ecosystems. These hominins utilised local and distant toolstone raw materials, indicating their ability to adapt to diverse ecological and elevation gradients. Through comparison of Ararat-1 with other sequences in the region, we highlight the spatia

Short-term occupations at high elevation during the Middle Paleolithic at Kalavan 2 (Republic of Armenia)

The Armenian highlands encompasses rugged and environmentally diverse landscapes and is characterized by a mosaic of distinct ecological niches and large temperature gradients. Strong seasonal fluctuations in resource availability along topographic gradients likely prompted Pleistocene hominin groups to adapt by adjusting their mobility strategies. However, the role that elevated landscapes played in hunter-gatherer settlement systems during the Late Pleistocene (Middle Palaeolithic [MP]) remains poorly understood. At 1640 m above sea level, the MP site of Kalavan 2 (Armenia) is ideally positioned for testing hypotheses involving elevation-dependent seasonal mobility and subsistence strategies. Renewed excavations at Kalavan 2 exposed three main occupation horizons and ten additional low densities lithic and faunal assemblages. The results provide a new chronological, stratigraphical, and paleoenvironmental framework for hominin behaviors between ca. 60 to 45 ka. The evidence presented suggests that the stratified occupations at Kalavan 2 locale were repeated ephemerally most likely related to hunting in a high-elevation within the mountainous steppe landscape.info:eu-repo/semantics/publishedVersio

Pharmacological modulation of ethanol reinforcing properties in mice exposed to a new animal model of post-traumatic stress disorder

Trabajo presentado al Seminario de Unidad Neurobiología Molecular y Neuropatología del Instituto de Neurociencias, celebrado online el 20 de julio de 2021.Peer reviewe

Efectos del cannabidiol en un nuevo modelo animal de trastorno de estrés postraumático

En esta tesis doctoral se ha desarrollado un nuevo modelo animal de trastorno de estrés postraumático (TEPT) que induce cambios pronunciados sobre el condicionamiento al miedo y la ansiedad, que se mantienen hasta 9 semanas después de haber finalizado la fase de inducción (Artículo 1). Estas modificaciones se acompañan de alteraciones en la expresión génica de la hormona liberadora de corticotropina (CRF), receptores cannabinoides 1 (CNR1) y 2 (CNR2) y transportador de serotonina (5HTT) en distintos núcleos cerebrales. Debido a la larga duración de las características conductuales y de cambios funcionales cerebrales, el modelo ha permitido la evaluación de tratamientos farmacológicos de varias semanas de duración. Se ha observado que la administración de cannabidiol (CBD) o sertralina (STR) regula las alteraciones producidas por el modelo, si bien se consigue un efecto sinérgico o aditivo con la combinación de ambos fármacos (Artículo 1).Por otra parte, la larga duración de las alteraciones inducidas por el modelo ha permitido su combinación con los procedimientos experimentales del consumo voluntario (CV) y autoadministración oral de etanol (AAO). Los roedores expuestos al modelo animal de TEPT presentan una mayor vulnerabilidad por los efectos reforzantes y motivacionales del alcohol, simulando de esta manera el consumo problemático de alcohol en pacientes con TEPT. Estas conductas se acompañan de alteraciones en la expresión génica de la tirosina hidroxilasa (TH), receptor mu-opioide (Opmr1) y 5HTT en distintos núcleos cerebrales, indicando la implicación de los sistemas dopaminérgico, opioide y serotoninérgico en el desarrollo de la conducta. La administración de STR y naltrexona (NTX) reduce el consumo y la motivación por obtener alcohol en ratones expuestos al modelo animal de TEPT y regula los cambios en la expresión génica de las distintas dianas evaluadas (Artículo 2).En conjunto, los datos obtenidos en la presente tesis doctoral ponen de manifiesto que: i) el nuevo modelo animal crónico de TEPT desarrollado produce alteraciones en la respuesta emocional que se mantienen a largo plazo, incluyendo un mayor rasgo de ansiedad, problemas en la extinción de memorias aversivas (miedo), y una elevada vulnerabilidad por la acción reforzante y motivacional del alcohol, así como cambios significativos en la expresión génica de distintas dianas, ii) la combinación farmacológica de CBD y STR puede resultar un tratamiento eficaz en pacientes con TEPT, iii) la combinación de la STR con una dosis subefectiva de NTX podría ser una herramienta a tener en cuenta en el tratamiento del trastorno dual TEPT y trastornos por uso de alcohol (TUA).Peer reviewe

Sertraline plus naltrexone reduced ethanol consumption and motivation in mice exposed to a chronic animal model of post-traumatic stress disorder

Trabajo presentado al 33rd ECNP Congress Virtual, celebrado del 12 al 15 de septiembre de 2020.[Background]: Post-traumatic stress disorder (PTSD) is a chronic and disabling mental illness often associated with other neuropsychiatric disorders such as substance abuse, being alcohol use disorder (AUD) the most prevalent in PTSD patients [1,2]. The development of animal models of the dual pathology PTSD-AUD is crucial to improve our understanding of the behavioural and neurobiological alterations involved to facilitate the development of new drugs.[Aims]: The main goals of this study were to develop a new animal model of PTSD-AUD dual pathology, and to evaluate the efficacy of sertraline, naltrexone or its combination to modulate ethanol consumption and motivation in C57BL/6J mice.[Methods]: C57BL/6J male mice were exposed to several stressful stimuli for 5 weeks, alternating 3 weeks of stress exposure with 2 weeks of resting, to induce the animal model of PTSD. Behavioural disturbances induced by the exposure to the PTSD model were evaluated by the fear conditioning paradigm (FC, week 6) and the novelty suppressed feeding test (NSFT, week 7). After the behavioural evaluation, a group of mice was exposed to the voluntary ethanol consumption paradigm (VC, weeks 8-12). Furthermore, another group of mice was exposed to the oral ethanol self-administration paradigm (OEA, weeks 8-15) to evaluate the effects of sertraline, naltrexone or its combination on ethanol consumption and motivation. Finally, relative gene expression analyses of mu-opioid receptor (Opmr1) in the nucleus accumbens, tyrosine hydroxylase (TH) in the ventral tegmental area and serotonin transporter (5HTT) in the dorsal raphe nucleus were carried out.[Results]: Mice exposed to the animal model of PTSD showed increased freezing time in the FC paradigm (Student's t-test, P<0.001), and enhanced latency time and reduced food consumption compared with controls in the NSFT (Student's t-test, P<0.05). In addition, the exposure to the animal model of PTSD significantly increased ethanol consumption and preference in the VC paradigm (Two-way RM ANOVA, P<0.001), and enhanced ethanol consumption and motivation in the OEA paradigm (Two-way RM ANOVA, P<0.001). Interestingly, the administration of sertraline plus naltrexone in the OEA paradigm led to a synergistic effect, significantly reducing ethanol consumption and motivation (Two-way RM ANOVA, P<0.001). Real time PCR studies showed reduced relative gene expression of Opmr1 and TH while increased 5HTT gene expression in mice exposed to the animal model of PTSD-AUD compared with controls (Student's t-test, P<0.001). These changes were normalised with sertraline, naltrexone or sertraline plus naltrexone administration (Two-way ANOVA, P<0.01).[Conclusions]: The present study identify a new animal model of PTSD-AUD dual pathology characterised by a long-lasting vulnerability to ethanol reinforcing and motivational actions and associated gene expression disturbances. Interestingly, the combination of sertraline plus naltrexone significantly reduced ethanol consumption and motivation, effects that were accompanied by the normalisation of relative gene expression alterations in Opmr1, TH and 5HTT. Future studies are warranted to evaluate the therapeutic potential of sertraline plus naltrexone combination in patients with PTSD-AUD dual pathology, and to further explore the neurobiological mechanisms involved.Peer reviewe

The administration of sertraline plus naltrexone reduces ethanol consumption and motivation in a long-lasting animal model of post-traumatic stress disorder

This study was aimed to evaluate the effects of sertraline (STR) and/or naltrexone (NTX) on ethanol consumption and motivation in an animal model of post-traumatic stress disorder (PTSD) and alcohol use disorder (AUD). Male C57BL/6J mice were submitted to an intermittent and progressively increasing stressful stimuli simulating PTSD behavioural features. Behavioural alterations were explored by the fear conditioning (FC), novelty suppressed feeding test (NSFT) and acoustic startle response (ASR) paradigms. Afterwards, mice were evaluated in the voluntary ethanol consumption (VC) and the oral ethanol self-administration (OEA) paradigms. The effects of STR (10 mg/kg) and/or NTX (0.7 mg/kg) on ethanol consumption and motivation were analysed in the OEA. Furthermore, relative gene expression analyses of tyrosine hydroxylase (Th), mu-opioid receptor (Oprm1) and 5-hydroxitryptamine transporter (Slc6a4) were performed in the ventral tegmental area (VTA), nucleus accumbens (NAcc) and dorsal raphe nucleus (DR), respectively. PTSD-like mice presented increased fear-related memory, anxiety-like behaviours, and startle response, as well as enhanced ethanol consumption and motivation in the VC and OEA paradigms. Interestingly, STR plus NTX combination significantly reduced ethanol intake and motivation in the OEA. Gene expression analyses revealed reduced Th and Oprm1 whereas Slc6a4 gene expression increased in PTSD-like mice. STR and/or NTX modulated Th and Slc6a4 gene expression changes in PTSD-like mice. Furthermore, NTX increased Oprm1 gene expression revealing a synergistic action when combined with STR. These results provide evidence about the efficacy of the STR plus NTX to attenuate ethanol reinforcement and motivation in an animal model of PTSD and AUD dual pathology.This research was supported by ‘Instituto de Salud Carlos III’ (RETICS, RD12/0028/0019 and RD16/0017/0014), ‘Plan Nacional Sobre Drogas’ (PNSD 2016I016) and ‘Ministerio de Economía y Competitividad’ (FIS, PI14/00438) to J.M.Peer reviewe

Cannabidiol and sertraline regulate behavioral and brain gene expression alterations in an animal model of PTSD

This study evaluated the effects of cannabidiol (CBD) and/or sertraline (STR) on behavioral and gene expression alterations induced by a new chronic animal model of post-traumatic stress disorder (PTSD). C57BL/6J male mice were repeatedly exposed to physical and psychogenic alternate stressful stimuli. Fear-related memory and anxiety-like behaviors were evaluated. The effects of the administration of CBD (20 mg/kg, i.p.) and/or STR (10 mg/kg, p.o.) were analyzed on behavioral and gene expression changes induced by the model of PTSD. Gene expression alterations of targets related with stress regulation, endocannabinoid and serotonergic systems were analyzed by real-time PCR. The results revealed an increased and long-lasting fear-related memory and anxiety-like behaviors in mice exposed to the animal model of PTSD. Treatment with CBD improved these behaviors in PTSD animals, effects that were significantly potentiated when combined with STR. Gene expression analyses revealed a long-term increase of corticotropin releasing factor (Crf) that was significantly normalized with the combination CBD plus STR. Cannabinoid receptors (Cnr1 and Cnr2) were up regulated in PTSD mice whereas the serotonin transporter (Slc6a4) was reduced. Interestingly, CBD and STR alone or combined induced a significant and marked increase of Slc6a4 gene expression. These results point out the cooperative action of the combination CBD plus STR to enhance fear extinction and reduce anxiety-like behaviors, normalizing gene expression alterations in this animal model of PTSD and suggesting that the combination of CBD with STR deserves to be further explored for the treatment of patients with PTSD.This work was supported by “Instituto de Salud Carlos III” (RETICS, RD12/0028/0019 and RD16/0017/0014), “Plan Nacional Sobre Drogas” (PNSD, 2016I016 and 2019I012) and “Ministerio de Economía y Competitividad” (FIS, PI14/00438 and PI18/00576) to JM. We also acknowledge financial support from the Spanish Ministerio de Economía y Competitividad from the “Severo Ochoa” Programme for Centres of Excellence in R&D (SEV-2017-0723).Peer reviewe